This invention concerns a process for the determination of cells secreting cellular contents such as proteins and/or glycoproteins, hormones, or growth factors, from a medium containing these cells and cells of the same species or genus. The invention also concerns a device for implementing this process.
In a number of applications in the biological, pharmacological, and medical field, there is a need to be able to differentiate cells secreting cellular substances from cells of the same species and genus. Thus, for example, it is of considerable importance for the formation of monoclonal antibodies to determine stimulated lymphocytes from a suspension of stimulated lymphocytes and unstimulated lymphocytes.
Certain lymphocytes form antibodies (glycoproteins) against foreign substances in the organism, such as a foreign protein, which is injected into the bloodstream. If such lymphocytes stimulated for the formation and release of antibodies are fused with a tumor cell, such as a myeloma cell, there is a chance that a so-called hybidoma cell will be formed which has the features of both parent components. This cell produces antibodies, specifically only against the foreign substance involved (so-called monoclonal antibodies). It is practically immortal and can multiply permanently in nutrient media, in contrast to a normal differentiated cell such as the lymphocyte.
However, since only a very small number of the many lymphocytes found in the lymphatic system are stimulated, and since only the stimulated lymphocytes should be supplied for fusion if possible, the stimulated lymphocytes must be separated from the unstimulated lymphocytes in a selection process. However, there are difficulties here even in recognizing the stimulated lymphocytes, since they cannot be differentiated from the unstimulated lymphocytes by microscopic observation.
As another example, it might also be mentioned that some microorganisms are able to lyse living yeast cultures by secreting certain cellular substances (primarily glycoproteins). These toxins are called "killer factors". "Killer yeasts" can cause difficulties in the production of beer. Even with a small degree of contamination, they produce changes in the fermentation and the quality of the beer. The existence and consequences of killer reactions have been established in brewer's yeasts, wine yeasts, baking yeasts, and also with other species of cells, such as Hansenula, Pichia, Deberyomyces, Kluyveromyces, Candida, and Torulopsis, for example.
In the cases mentioned, and in many others, it is first a question of recognizing the secreting cells in order to be able to initiate other measures. These other measures can consist of then segregating these cells, as in the case of the stimulated lymphocytes, for example. In the case of killer yeast cells, the recognition of such cells is useful initially for the monitoring of the fermentation process. This in turn can lead to the introduction of measures to avoid the contamination.
It is therefore a principal object of the present invention to provide a process which makes it possible to determine cells secreting cellular substances such as proteins and glycoproteins, hormones, or growth factors from a suspension containing cells of the same species and genus.
It is also an object of this invention to provide a device for implementing the process.